The Effect Of NSE Gene Interference On Luteal Cell Proliferation And The Expression Of Its Endocrine Markers

Researches have confirmed that neuron specificity enolase (NSE), synapothysin (SYP),5-hydroxytryptamine (5-HT) and S100B protein (S100B), the four kinds of diffuseneuroendocrine cell markers, are expressed in corpus luteum cells of goat. However, the rolesof these four substances in the corpus luteum and their interrelations are still not clear. In thepresent experiment, proliferation of immortalized corpus luteum cells of goats and the fourdiffuse neuroendocrine cell markers were detected before and after interference through cellculture, RNA interference, MTT method, immunocytochemistry, immunofluorescencecytochemistry and real-time fluorescent quantitative PCR. The main results are described asfollows:1、MTT results showed that after the interference proliferation of immortalized corpusluteum cells is restrained.2、Immunocytochemistry showed that the expression of NSE in negative control groupwas higher than interference group.After interfering RNA in immortalized corpus luteum cells,some cells were observed without the expression of NSE. SYP and S100B were observedlittle expression both in negative control group and interference group. Also,5-HT did notchange significantly before and after the interference.3、Immunofluorescence cytochemistry indicated that NSE expressed in immortalizedcorpus luteum cells before and after interference, but largely expressed in the negative corpusluteum cells.5-HT expressed in cytoplasm, and lower than NSE. Only a few cells expressed5-HT but with a high expression after interfering the corpus luteum cells. SYP and S100Bwere observed little expression without significant change both in negative control group andinterference group.4. Real-time fluorescent quantitative PCR results showed that after interfering NSE themRNA expression levels were significantly lower than the negative control group inimmortalized corpus luteum cells (P <0.05). SYP and S100B were observed little expressionwithout significant change both in negative control group and interference group.The results show that corpus luteum cells growth is restrained after interfering with NSEgene, which suggests that NSE may have the effects of support and nutrition of un-glycolysison corpus luteum cells Its expression promote the proliferation of orpus luteum cells. knocking out of NSE genes can resulting in abnormal expression of5-HT in individual cells,while, the expression of SYP and S100B have no obvious change, Giving a suggest that thereis a relationship between the expression of5-HT and NSE, S100B and SYP may played asupporting role in corpus luteum.